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Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A
The 5’ cap of mRNA plays a critical role in mRNA processing, quality control and turnover. Enzymatic availability of the 5’ cap governs translation and could be a tool to investigate cell fate decisions and protein functions or develop protein replacement therapies. We have previously reported on th...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10108117/ https://www.ncbi.nlm.nih.gov/pubmed/36408753 http://dx.doi.org/10.1002/cbic.202200522 |
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author | Klöcker, Nils Anhäuser, Lea Rentmeister, Andrea |
author_facet | Klöcker, Nils Anhäuser, Lea Rentmeister, Andrea |
author_sort | Klöcker, Nils |
collection | PubMed |
description | The 5’ cap of mRNA plays a critical role in mRNA processing, quality control and turnover. Enzymatic availability of the 5’ cap governs translation and could be a tool to investigate cell fate decisions and protein functions or develop protein replacement therapies. We have previously reported on the chemical synthesis of 5’ cap analogues with photocleavable groups for this purpose. However, the synthesis is complex and post‐synthetic enzymatic installation may make the technique more applicable to biological researchers. Common 5’ cap analogues, like the cap 0, are commercially available and routinely used for in vitro transcription. Here, we report a facile enzymatic approach to attach photocleavable groups site‐specifically to the N(2) position of m(7)G of the 5’ cap. By expanding the substrate scope of the methyltransferase variant GlaTgs V34A and using synthetic co‐substrate analogues, we could enzymatically photocage the 5’ cap and recover it after irradiation. |
format | Online Article Text |
id | pubmed-10108117 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-101081172023-04-18 Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A Klöcker, Nils Anhäuser, Lea Rentmeister, Andrea Chembiochem Research Articles The 5’ cap of mRNA plays a critical role in mRNA processing, quality control and turnover. Enzymatic availability of the 5’ cap governs translation and could be a tool to investigate cell fate decisions and protein functions or develop protein replacement therapies. We have previously reported on the chemical synthesis of 5’ cap analogues with photocleavable groups for this purpose. However, the synthesis is complex and post‐synthetic enzymatic installation may make the technique more applicable to biological researchers. Common 5’ cap analogues, like the cap 0, are commercially available and routinely used for in vitro transcription. Here, we report a facile enzymatic approach to attach photocleavable groups site‐specifically to the N(2) position of m(7)G of the 5’ cap. By expanding the substrate scope of the methyltransferase variant GlaTgs V34A and using synthetic co‐substrate analogues, we could enzymatically photocage the 5’ cap and recover it after irradiation. John Wiley and Sons Inc. 2022-12-08 2023-01-17 /pmc/articles/PMC10108117/ /pubmed/36408753 http://dx.doi.org/10.1002/cbic.202200522 Text en © 2022 The Authors. ChemBioChem published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Research Articles Klöcker, Nils Anhäuser, Lea Rentmeister, Andrea Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A |
title | Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A |
title_full | Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A |
title_fullStr | Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A |
title_full_unstemmed | Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A |
title_short | Enzymatic Modification of the 5’ Cap with Photocleavable ONB‐Derivatives Using GlaTgs V34A |
title_sort | enzymatic modification of the 5’ cap with photocleavable onb‐derivatives using glatgs v34a |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10108117/ https://www.ncbi.nlm.nih.gov/pubmed/36408753 http://dx.doi.org/10.1002/cbic.202200522 |
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